Persistent Changes in T Cell Receptor Surface Organization Visualized by Quantum Dot Probes with k-Space Image Correlation Spectroscopy (50.2)
| Autor: | Karlo Perica, Joan Bieler, Yu Li, Aaron Selya, Michael Edidin, Jonathan Schneck |
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| Rok vydání: | 2010 |
| Předmět: | |
| Zdroj: | The Journal of Immunology. 184:50.2-50.2 |
| ISSN: | 1550-6606 0022-1767 |
| DOI: | 10.4049/jimmunol.184.supp.50.2 |
| Popis: | There is growing evidence that the surface pattern and spatial distribution of T cell receptors (TCRs) on cell surfaces plays a critical role in T Cell antigen detection and effector function, from micron-sized Supramolecular Adhesion Clusters to microclusters 100’s of nanometers in diameter. To date, TCR organization patterns have been primarily examined in the context of antigen stimulation or immediately thereafter, on a timescale of seconds to minutes, despite evidence from binding studies that shows changes persist many days post-activation. Here, we present a method for imaging long-term TCR lateral surface organization on live cells at two relevant length scales: 10’s and 100’s of nms. TCR are labeled with monomeric biotin-MHC/peptide or antibody fragments, followed by streptavidin-quantum dots (QD). We have found that QD “blinking” is a useful parameter for detecting TCR organization on a nm scale, since electronic coupling between blinking states of adjacent QDs reports single or small clusters of receptors. Simultaneously, we quantify organization at the scale of 100’s of nms by epifluorescense. As proof of principle, we have demonstrated that in vitro activated T cells have enhanced clustering at the 10’s and 100’s nm scales compared to naïve counterparts, supporting results from MHC dimer binding assays. Clustering can also be modulated by IL-2 signaling for active, but not naïve, T cells. |
| Databáze: | OpenAIRE |
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